2,3-epoxy alcohols, acids and derivatives as anti retroviral chemotherapeutic agents

ABSTRACT

The present invention is related to compounds, compositions and methods of treating viral infections. Compounds of the present invention have the following general formula: ##STR1## wherein R is selected from --CH 2  OH, --CO 2  R 2 , --CONR 3  R 4 , or COR 5 , wherein R 2  is hydrogen or a lower alkyl group, R 3  and R 4  are each independently hydrogen or a lower alkyl group, R 5  is an amino acid residue bound via a terminal nitrogen or peptide having at least two amino acid residues; and wherein R 1  is C 5  -C 13  alkyl, aryl, aralkyl, aralkyl(lower alkyl)ether, or C 5  -C 13  alkyl(lower alkyl)ether.

This application is a divisional of application Ser. No. 07/286,977,filed on Dec. 20, 1988 now U.S. Pat. No. 5,190,969, the entire contentsof which are hereby incorporated by reference.

BACKGROUND OF THE INVENTION

Retroviruses have long been associated with neoplastic disease in avianand mammalian species. The discovery of infectious human retroviruseswhich are associated with malignancies has spawned a surge of interestin these agents. Recent work linking a retrovirus, HumanImmunodeficiency Virus (HIV), as an etiologic agent of Acquired-ImmuneDeficiency Syndrome (AIDS) (Gallo, R. C. (1987) Sci. Amer., 256, 47-56)has further intensified this interest.

Research on antiviral agents has progressed far in the past decade inresponse to the need to treat infections of HIV. The majority of thework on antiretrovirals has focused on the use of nucleoside derivativesas inhibitors of the retroviral reverse transcriptase (Fischl, M. A., etal., N. Eng. J. Med., 317, 185-191 (1987);J. E. Dahlberg, H. Mitsuya, S.B. Blum, S. Broder and S. A. Aaronson, Proc. Natl. Acad. Sci., 84,2469-2473 (1987); S. Broder, in "Human Retroviruses, Cancer, and AIDS",D. Bolognes; ed., Alan R. Liss, Inc., NY, 1988, pp. 365-380; M. S. Hirshand J. C. Kaplan, Antimic Agents and Chemotherapy, 31, 839-843 (1987).Many of these agents have characteristics which restrict their clinicalutility (D. D. Richman, et al., N. Eng. J. Med., 317, 192-197 (1987);Terasaki, T., Pardridge, W. M., J. Infect. Disease, 158, 630-632(1988)).

Viral proteases are one of the enzymes which are essential components ofvirion assembly (J. Wellink and A. Van Kammen, Arch. Virol., 98, 1-26(1988)). Site specific mutagenesis of the presumed active site ofretroviral proteases has demonstrated that active enzyme is necessaryfor the production of mature, infectious virus particles (I. Katoh, etal, Virology, 145, 280-292 (1985); N. E. Kohl, et al. Proc. Natl. Acad.Sci., 85, 4686-4690 (1988); S. Seelmeier, H. Schmidt, V. Tura, and K.von der Helm, Proc. Natl. Acad. Sci., 85, 6612-6616 (1988)).Consequently, interference of proteolytic processing through the use ofenzyme inhibitors is an attractive chemotherapeutic target for thetreatment of viral diseases. Expression of HIV protease in E. coli (S.Seelmeier, H. Schmidt, V. Tura, and K. von der Helm, Proc. Natl. Acad.Sci., 85, 6612-6616 (1988); M. C. Graves, J. J. Lim, E. P. Heimer and R.A. Kramer, Proc. Natl. Acad. Sci., 85, 2449-2453 (1988); C. DeBouck, etal., Proc. Natl. Acad. Sci., 84, 8903-8906 (1987); J. Mous, E. P. Heimerand S. F. J. Le Grice, J. Virol., 62, 1433-1436 (1988)), and chemicalsynthesis (T. D. Copeland and S. Oroszlan, Gene Anal. Tech., 5, 109-115(1988); J. Schneider and S. B. H. Kent, Cell, 54, 363-368 (1988); R. F.Nutt, et al, Proc. Natl. Acad. Sci., 85, 7129-7133 (1988)) of activeenzyme have provided in vitro systems to facilitate the study of thisprotein.

The antifungal antibiotic cerulenin,4-Oxo-2R,3S-epoxy-trans,trans-2,5-dodecadienyl amide has been reportedto exhibit antiretroviral activity against Rous Sarcoma Virus (H.Goldfine, J. B. Harley and J. A. Wyke, Biochem. Biophys. Acad., 512,229-240 (1978)) and Murine Leukemia Virus (I. Katoh, Y. Yoshinaka and R.B. Luftig, Virus Res., 5, 265-276 (1986)) in vitro. Originally studiedfor its inhibition of fatty acid synthesis through the inhibition ofβ-ketoacetyl transferase, (for review, see S. Omura, BacteriologicalRev., 40, 681-697 (1976)) cerulenin apparently interferes withpolypeptide processing during virus particle assembly (K. Ikuta and R.B. Luftig, Virology, 154, 195-206 (1986)). It has been uncertain at whatstage of processing the inhibition was occurring, but interference withenzyme catalyzed proteolytic cleavage of virus precursor polypeptideshas been assumed to be likely. Cerulenin has also been recently reportedto inhibit viral polyprotein processing in HIV infected cells (Pal, R.,et al (1988), Proc. Natl. Acad. Sci., 85, 9283-9286). Since cerulenin isa potent inhibitor of fatty acid synthesis, it exhibits a relativelyhigh toxicity (Matsumae, J. Antibiotic., 17a, 1 (1964); Pal, R. et al(1988)). The present invention has been discovered with the abovebackground and disadvantages in mind.

SUMMARY OF THE INVENTION

The present invention is related to compounds useful in treating viralinfections. The compounds of the present invention have the followinggeneral formula: ##STR2## wherein R is selected from --CH₂ OH, --CO₂ R²,--CONR³ R⁴ or COR⁵, wherein R² is hydrogen or a lower alkyl group, R³and R⁴ are each independently hydrogen or a lower alkyl group, R⁵ is anamino acid residue bound via a terminal nitrogen on said amino acid or apeptide having at least two amino acid residues; and wherein R¹ isaralkyl, aralkyl(lower alkyl)ether or C₅ -C₁₃ alkyl(lower alkyl)ether.

The compound of the present invention also includes compounds having thegeneral formula: ##STR3## wherein R⁵ is an amino acid residue bound viaa terminal nitrogen on said amino acid or a peptide having at least twoamino acid residues; and wherein R¹ is C₅ -C₁₃ alkyl.

The present invention is also directed to compositions comprising acompound having the formula: ##STR4## wherein R is selected from --CH₂OH, --CO₂ R², --CONR³ R⁴, or COR⁵, wherein R² is hydrogen or a loweralkyl group, R³ and R⁴ are each independently hydrogen or a lower alkylgroup, R⁵ is an amino acid residue bound via a terminal nitrogen on saidamino acid, or a peptide having at least two amino acid residues; andwherein R¹ is aralkyl, aralkyl(lower alkyl)ether or C₅ -C₁₃ alkyl(loweralkyl)ether; and a pharmaceutically acceptable carrier, as well ascompositions comprising a compound having the formula: ##STR5## whereinR⁵ is an amino acid residue bound via a terminal nitrogen on said aminoacid, or a peptide having at least two amino acid residues; and whereinR¹ is C₅ -C₁₃ alkyl, and a pharmaceutically acceptable carrier.

Further, the present invention is directed to a method of treating viralinfections to a host in need thereof by administering an anti-viraleffective amount of the compound having the formula: ##STR6## wherein Ris selected from --CH₂ OH, --CO₂ R², --CONR³ R⁴, or COR⁵, wherein R² ishydrogen or a lower alkyl group, R³ and R⁴ are each independentlyhydrogen or a lower alkyl group, R⁵ is an amino acid residue bound via aterminal nitrogen or peptide having at least two amino acid residues;and wherein R¹ is C₅ -C₁₃ alkyl, aryl, aralkyl, aralkyl(loweralkyl)ether, or C₅ -C₁₃ alkyl(lower alkyl)ether. By host is meantmembers from the classes of mammals or aves, or any animal in need ofanti viral therapy.

By lower alkyl is meant C₁ -C₄, straight, and branched hydrocarbonchains.

By aryl is meant an organic radical derived from an aromatic hydrocarbonby removal of one atom. An example of an aryl is a phenyl group.

By aralkyl is meant an arylated alkyl wherein an alkyl H atom issubstituted by an aryl group. The alkyl group can be a straight ofbranched chain having 1-6 carbon atoms.

By amino acid residue is meant any of the naturally occurring orsynthetic amino acids commonly found or synthesized. Examples of thenaturally occurring amino acids are glycine, alanine, valine, leucine,isoleucine, methionine, proline, pheylalanine, tryptophan, serine,threonine, cysteine, tyrosine, asparginine, glutamine, aspartic acid,glutamic acid, lysine, arginine, histidine and the like.

By alkyl is meant an alkyl group having an alkyl chain length of from 5to 13 carbon atoms.

By aralkyl (lower alkyl) ether is meant a aralkyl group attached via anether linkage to a lower alkyl chain having from 1 to 4 carbon atoms.

Of the viral infections that can be treated, any virus, which encodesfor a protease, could be treated with the compounds of the presentinvention. Retroviral infections can be treated with the compounds ofthe present invention. Examples of such viruses are Type C, Type Dretroviruses, HTLV-1, HTLV-2, HIV-1, HIV-2, murine leukemia virus,murine mammary tumor virus, feline leukemia virus, bovine leukemiavirus, equine infectious anemia virus, avian sarcoma viruses such asrous sarcoma virus, and the like.

These and other advantages achieved by the present invention will becomeapparent upon reading of the detailed description of the PreferredEmbodiments.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Generally, the process of the present invention involves the asymmetricepoxidation of the allylic alcohol by the method of Gao et al. (J. Am.Chem. Soc., 1987, 109 5765-80) with (+)-diisopropyl tartrate (DIPT),tert-butylhydroperoxide (TBHP), and titanium tetraisopropoxide in thepresence of 4A powdered molecular sieves to produce the 2S-epoxyalcohol. The allylic-alcohol is obtained by 1) metallation of theacetylene followed by reaction with formaldehyde to afford a propargylalcohol which is selectively hydrogenated with Lindlars catalyst or 2)metallation of the acetylene followed by reaction with formaldehyde toafford a propargyl alcohol which is partially reduced with sodiumbis(2-methoxyethoxy) aluminum hydride or 3) monoalkylation ofcis-2-butene-1,4-diol. The epoxy alcohol may be oxidized by 1) RuCl₃ /H₅IO₆ or 2) pyridinium dichromate in DMF, to yield the epoxy acid.Esterification of the acid with CH₂ N₂ followed by treatment with NH₄OH/MeOH affords the epoxy amide. Alternatively, the acid may be used toacylare a variety of nucleophilic substrates via its mixed anhydride byreaction of the epoxy acid with iso-butyl chloroformate andtriethylamine in THF followed by nucleophile (NH₄ OH, RNH₂, R₂ NH, erc).Epoxidation with (-)-diisopropyl tartrate rather than (+)-diisopropyltartrate in the above reaction scheme will afford the opposite opticalisomer.

The following examples, which are directed to the process of preparingspecific compounds, is in no way to be construed as limiting theinventive scope of the present invention. All percentages are by weightunless expressly stated to the contrary. Where necessary, the source ofthe starting materials are identified.

Experimental

All solvents were reagent grade and were used as received unless notedotherwise. Tetrahydrofuran was freshly distilled from benzophenone ketylprior to use. Dichloromethane was dried over 3A molecular sievesovernight. Ti(OiPr)₄ and tartrate esters were obtained from AldrichChem., distilled under reduced pressure and stored under argon prior touse. Triethylamine was distilled from CaC₂. Acetylenic substrates,2-decyn-1-ol, and cis-2-decene were obtained from either Farchan Labs orWiley Chem. and were used as received. Aqueous tert-butylhydroperoxidewas obtained from Aldrich and the isooctane solution prepared asdescribed by Gao et al. (J. Am. Chem. Soc., 1987, 109, 5765-80). Allother reagents were obtained from Aldrich Chemicals and were used asreceived. Flash chromatography was performed as described by Still etal. (J. Org. Chem. 1978, 43, 2923-25). NMR spectra were obtained on aVarian XL-200 or a Nicolet NT-300 in CDCl₃ and are reported in ppmdownfield from TMS. Mass spectra were obtained on a VG-Micromass ZAB-2Fmass spectrometer. Melting points and boiling points are uncorrected.

EXAMPLE 1 Preparation of 2-Dodecyn-1-ol

An anhydrous solution of 49.5 g (325 mmole) of 1-undecyne in 600 ml ofether was prepared under argon and cooled to -20° C. A solution ofn-BuLi in hexanes (130 ml, 2.5 M, 325 mmole) was then added to theacetylene solution dropwise over about 1.5 H. By the end of the additiona heavy white precipitate has formed. The cooling bath was removed andthe solution was stirred for an additional hour. Paraformaldehyde (10.76g, 358 mmole) was then added in one portion and the mixture was allowedto stir at room temperature under argon overnight. Saturated NH₄ Cl (400ml) was then added cautiously to the mixture with vigorous stirring andcooling as necessary. The ether layer was separated and washed twicewith 200 ml of water and 200 ml of brine, dried over MgSO₄, filtered,and the solvent evaporated to afford a cloudy oil. Distillation affords49.62 g of a clear oil, bp 95-105° C.(0.1 mm), which solidified uponstanding. ¹ H NMRS δ 4.25 (m, 2), 2.21 (tt, 2, J=2, 7 Hz), 1.6-1.1 (m,14), 0.88 (t, 3, J=7 Hz).

EXAMPLE 2 Preparation of 2-Tetradecyn-1-ol

2-Tetradecyn-1-ol was prepared as described above for 2-dodecyn-1-olexcept that 5.0 g (27 mmole) of 1-tridecyne in 200 ml of ether, 11.6 ml(29 mmole) of n-BuLi, and 874 mg (29 mmole) of paraformaldehyde wasused. The reaction afforded a white solid after workup which wascrystallized from petroleum ether to yield 4.78 g of white crystals, mp44-6° C. ¹ H NMR δ 4.25 (m, 2), 2.20 (m, 2), 1.6-1.2 (m, 20), 0.88 (t,3, J=7 Hz).

EXAMPLE 3 Preparation of cis-2-Decen-1-ol

To a solution of 10.0 g of 2-decyn-1-ol in 150 ml of petroleum ether wasadded 500 mg of Pd on CaCO₃ poisoned with lead (Lindlar catalyst).Quinoline (2.0 ml) was then added to this mixture and the flask wasevacuated (house vacuum, ca. 50 mm Hg) and flushed with H₂ three timesand left under a H₂ atmosphere. The mixture was stirred vigorously untiluptake of H₂ was no longer evident and TLC (SiO₂ /20% ether:pet. ether)showed no starting material. The catalyst was removed by filtrationthrough a 4 cm pad of celite and the filtrate was washed two times with100 ml of 1 N HCl, once with 100 ml of saturated NaHCO₃, once with 100ml of brine, dried over MgSO₄, filtered and the solvent evaporated togive a pale yellow oil. Kugelrohr distillation (100-120° C. oven, 0.2mm) affords 8.81 g of clear oil. ¹ H NMR δ 5.57 (m,2), 4.20 (t, 2, J=5Hz), 2.07 (dd, 2, J=6, 12 Hz), 1.5-1.1 (m, 10), 0.88 (t, 3, J=7 Hz).

EXAMPLE 4 Preparation of cis-2-Dodecen-1-ol

cis-2-Dodecen-1-ol was prepared as described above for cis-2-decen-1-olexcept that 35 g of 2-dodecyn-1-ol, 1.0 g of catalyst, and 3.0 ml ofquinoline was used. A clear oil, 33.86 g, was obtained after kugelrohrdistillation. ¹ H NMR δ 5.57 (m, 2), 4.19 (d, 2, J=5 Hz), 2.07 (dd, 2,J=6, 12 Hz), 1.5-1.2 (m, 14), 0.88 (t, 3, J=7 Hz).

EXAMPLE 5 Preparation of cis-2-Tetradecen-1-ol

cis-2-Tetradecen-1-ol was prepared as described above forcis-2-decen-1-ol except that 1.0 g of 2-tetradecyn-1-ol, 50 mg ofcatalyst, and 0.10 ml of quinoline was used. A clear oil, 0.72 g, wasobtained after kugelrohr distillation. ¹ H NMR δ 5.57 (m, 2) 4.19 9d, 2,J=6 Hz), 2.07 (dd, 2, J=6, 13 Hz), 1.5-1.2 (m, 18), 0.88 (t, 3, J=7 Hz).

EXAMPLE 6 Preparation of 4-Benzyloxy-(cis)-2-buten-1-ol

4-Benzyloxy-(cis)-2-buten-1-ol was prepared by the method of Danishefskiet al. (J. Am. Chem. Soc. 1985, 107, 3891-8). A clear oil was obtainedafter kugelrohr distillation. ¹ H NMR δ 7.4-7.3 (m, 5), 5.79 (m, 2),4.53 (s, 2), 4.14 (m, 4), 1.85 (br, 1).

EXAMPLE 7 Preparation of 4-Heptyloxy-(cis)-2-buten-1-ol

A 50% oil dispersion of NaH (9.4 g) was washed three times with 20 ml ofdry THF under argon and the remaining oil-free NaH was suspended in 600ml of dry THF. cis-2-Butene-1,4-diol (50 ml, 46.7 g, 530 mmole) was thenadded slowly by syringe to the suspension. After a small amount of diolwas added a gelatinous precipitate began to form, most of which wentback into solution by the end of the addition. The mixture was allowedto stir for an additional hour and n-heptyliodide (30.4 ml, 42.0 g, 186mmole) was added in one portion. The reaction was allowed to stirovernight. Aqueous NH₄ Cl was added to the mixture followed by 600 ml ofether. The organic layer was separated, washed with 200 ml of NaHCO₃,200 ml of brine, dried over MgSO₄, filtered and the solvent evaporatedto yield a pale yellow oil. Distillation afforded 23.82 g, bp 106-10° C.(0.1 mm). ¹ H NMR δ 5.76 (m, 2), 4.20 (t, 2, J=6 Hz), 4.04 (d, 2, J=6Hz), 3.44 (t, 2, J=7 hz), 2.22 (t, 1, J=6 Hz), 1.59 (br t, 2, J=7 hz)1.29 (br, 8), 0.88 (t, 3, J=7 hz).

EXAMPLE 8 Preparation of trans-2-Decen-1-ol

trans-2-Decen-1-ol was prepared by reduction of the propargyl alcohol bythe method described by Jones and Denmark (Org. Syn., 1985, 64, 182-8)using 15 g (97 mmole) of 2-decyn-1-ol in 40 ml of ether and 48 ml (162mmole) of Red-Al (sodium bis(2-methoxyethoxy)aluminum hybride) in 50 mlof ether. Kugelrohr distillation afforded 11.59 g of a clear oil. ¹ HNMR δ 5.66 (m, 2), 4.07 (d, 2, J=5 Hz), 20.3 (dd, 2, J=6, 13 hz), 1.68(br, 1), 1.4-1.2 (m, 10), 0.88 (t, 3, J=7 Hz).

EXAMPLE 9 Preparation of trans-2-Dodecen-1-ol

trans-2-Dodecen-1-ol was prepared by reduction of the propargyl alcoholby the method described by Jones and Denmark (Org. Syn., 1985, 64,182-8) using 6.0 g (33 mmole) of 2-dodecyn-1-ol in 25 ml of ether and16.2 ml of Red-Al in 30 ml of ether. Kugelrohr distillation afforded5.73 g of a clear oil. ¹ H NMR δ 5.66 (m, 2), 4.07 (d, 2, J=4 Hz), 2.03(dd, 2, J=6, 13 Hz), 1.62 (br, 1), 1.4-1.2 (m, 14), 0.88 (t, 3, J=7 Hz).

EXAMPLE 10 Preparation of trans-2-Tetradecen-1-ol

trans-2-Tetradecen-1-ol was prepared by reduction of the propargylalcohol by the method described by Jones and Denmark (Org. Syn., 1985,64, 182-8) using 8.0 g (38 mmole) of 2-tetradecyn-1-ol in 30 ml of etherand 16 ml (55 mmole) of Red-Al in 40 ml of ether. Kugelrohr distillationafforded 7.67 g of clear oil. ¹ H NMR δ 5.66 (m, 2), 4.08 (d, 2, J=5 Hz)2.03 (dd, 2, J=6, 13 Hz), 1.60 (br, 1), 1.4-1.2 (m, 18), 0.88 (t, 3, J=7Hz).

EXAMPLE 11 Preparation of (2S-cis)-3-Nonyloxirane Methanol

To a slurry of 500 mg of powdered 4A molecular sieves in 30 ml of dryCH₂ Cl₂ at 0° C. under argon, was added 232 mg (0.82 mmole) ofTi(Oi-Pr)₄ followed by 266 mg (1.14 mmole) of L(+)diisopropyl tartrate.The mixture was allowed to stir for 15 min. The mixture was cooled to-20° C. and a solution of 0.75 g (4.1 mmole) of cis-2-dodecene-1-ol in 5ml of CH₂ Cl₂, which has been dried over 3A molecular sieves for 15 min,was then added. After the catalyst had "aged" for 20 min, 2.15 ml of 3.8M (8.17 mmole) tert-butyl hydroperoxide in isooctane was added slowly bysyringe. The reaction was then allowed to stand in the freezer (-30° C.)for seven days. The mixture was then warmed to 0° C. and 5 ml of H₂ Owas added with stirring. After 30 min, 1.5 ml of 30% NaOH in saturatedbrine was added and the mixture was allowed to stir 60 min longer. Theorganic layer was separated and the aqueous layer was washed twice with20 ml of CH₂ Cl₂. The combined organic layers were dried over MgSO₄,filtered and evaporated to afford a sticky white solid.Recrystallization from Et₂ O:petroleum ether yielded 570 mg of aflocculent white solid mp 55-6° C. ¹ H NMR (CDCl₃) δ 3.86 (ddd, 1, J=4,8, 12 Hz), 3.7 (m, 1), 3.16 (dt, 1, J=4, 7 Hz) 3.0 (m, 1), 1.6-1.1 (m,12), 0.88 (t, 3, J=7 Hz). Mass spectra EI; M-31 (--CH₂ OH), Calc.169.1592, Found 169.1581.

EXAMPLE 12 Preparation of (2R-cis)-3-Nonyloxirane Methanol

The epoxidation was performed as described in Example 11 except thatD(-)diisopropyl tartrate was used. The reaction afforded 524 mg of awhite solid, mp 55-6° C. ¹ H NMR(CDCl₃) δ 3.86 (ddd, 1, J=4, 8, 12 Hz),3.7 (m, 1), 3.16 (dt, 1, J=4, 7 Hz), 3.0 (m, 1), 1.6-1.1 (m, 12), 0.88(t, 3, J=6 Hz). Mass Spectra; EI, M-31 (--CH₂ OH), Calc. 169.1592, Found169.1594.

EXAMPLE 13 Preparation of (2S-cis)-3-(Benzyloxymethyl)oxirane Methanol

To a slurry of 5.0 g of powdered 4A molecular sieves in 400 ml of dryCH₂ Cl₂ was added 2.09 g (8.42 mmole) of Ti(Oi-Pr)₄ and 2.76 g (11.8mmole) of L(+)diisopropyl tartrate at 0° C. under argon. The mixture wasallowed to stir for 10 min, cooled to -20° C., and 22.5 ml of 3.8 Mtert-butyl hydroperoxide in isooctane is added slowly at -20° C. Thecatalyst was allowed to "age" for 30 min. A solution of 7.50 g (42.1mmole) of Z-4-(benzyloxy)-2-buten-1-ol in 15 ml of CH₂ Cl₂ was driedover 3A molecular sieves for 15 min and then added to the mixture at-20° C. over 10 min. The mixture was allowed to stand in a refrigeratorat 2° C. for 8 days. The reaction mixture was removed from therefrigerator and 40 ml of H₂ O was added with stirring. After stirringfor 30 min, 15 ml of 30% NaOH in saturated brine was added and themixture was allowed to stir 60 min longer. The organic layer wasseparated and the aqueous layer washed twice with 50 ml of CH₂ Cl₂. Theorganic layers were combined, dried over MgSO₄, filtered and evaporatedto afford a milky, pale yellow oil. Distillation yielded 4.80 g of aclear viscous oil, bp 132-5° C. (0.2 mm Hg). ¹ H NMR, δ 7.4-7.2 (m, 5),4.58 (dd, 2, J=12, 20 Hz), 3.7 (m, 4), 3.27 (ddd, 2, J=5, 10, 15 Hz),1.98 (t, 1, J=6 Hz). Mass Spectra; EI; M⁺, Calc. 194.0942, Found194.0929.

EXAMPLE 14 Preparation of (2R-cis)-3-(Benzyloxymethyl)oxirane Methanol

The epoxidation was performed as described in Example 13 except with 8.0g of powdered 4A sieves, 600 ml CH₂ Cl2, 3.34 g (13.5 mmole) ofTi(OiPr)₄, 4.41 g (19.8 mmole) of D(-)diisopropyl tartrate, 35.5 ml of3.8 M tert-butylhydroperoxide in isooctane, and 11.98 g of(Z)-4-(benzyloxy)-2-butene-1-ol. Workup with 70 ml H₂ O and 25 mlNaOH/Brine afforded 8.65 g of clear oil after distillation, bp 132-4° C.(0.1 mm Hg). ¹ H NMR δ (CDCl₃) 7.4-7.2 (m, 5), 4.58 (dd, 2, J=12, 20Hz), 3.7 (m, 4), 3.27 (ddd, 2, J=5, 10, 15 Hz), 2.17 (t, 1, J=6 Hz).Mass spectra: EI, M⁺, Calc. 194.0942, Found 194.0960.

EXAMPLE 15 Preparation of (2S-cis)-3-Heptyloxirane Methanol

A suspension of 3.00 g of 4A powdered molecular sieves in 60 ml of dryCH₂ Cl₂ was cooled to 0° C. under argon. To the cooled suspension wasadded 1.09 g (3.80 mmole) of Ti(OiPr)₄ and 1.24 g (5.32 mmole) ofL(+)diisopropyl tartrate via syringe. The mixture was stirred for 15 minand cooled to -20° C. A solution of tert-butylhydroperoxide in isooctane(3.8 M, 7.57 ml, 28.8 mmole) was added slowly to the mixture thecatalyst allowed to "age" for 30 min at -20° C. A solution of 3.00 g(19.2 mmole) of Z-2-decen-1-ol in 10 ml of CH₂ Cl₂ was dried over 3Amolecular sieves for 15 min before being added to the mixture slowly,with stirring, at -20° C. The solution was then placed in the freezer(-30° C.) and allowed to stand for 4 days. The mixture was warmed to 0°C. and poured into a solution of 2.00 g of citric acid and 5.50 g ofFeSO₄ in 15 Ml H₂ O. The organic layer was separated and the aqueouslayer washed twice with 20 ml of CH₂ Cl₂. The CH₂ Cl₂ layers werecombined, dried over MgSO₄, filtered and evaporated to yield a stickywhite solid. Recrystallization from Et₂ O;petroleum ether afforded 1.78g of flocculent white solid, mp 42-3° C. ²⁵ -3.4' (c 1.1, CHCl₃); ¹ HNMR δ 3.86 (ddd, 1, J=4, 8, 12 Hz), 3.67 (ddd, 1, J=5, 7, 12 Hz), 3.16(dt, 1, J=4, 7 Hz), 3.03 (m, 1), 1.74 (m, 1) 1.6-1.2 (m, 12), 0.88 (t,3, J=6 Hz). Mass spectra: EI; M-31 (--CH₂ OH), Calc. 141.1279, Found141.1281.

EXAMPLE 16 Preparation of (2R-cis)-3-Heptyloxirane Methanol

The epoxidation was performed as described in Example 15 except thatD(-)diisopropyl tartrate was used. The reaction afforded 1.70 g offlocculent white crystals, mp 40-1° C. ¹ H NMR (CDCl₃) δ 3.86 (ddd, 1,J=4, 8, 12 Hz), 3.67 (ddd, 1, J=5, 7, 12 Hz), 3.16 (dt, 1, J=4, 7 Hz),3.03 (m, 1), 1.77 (dd, 1, J=5, 7 Hz), 1.6-1.2 (m, 12), 0.88 (t, 3, J=6Hz). Mass Spectra: EI, M-31 (--CH₂ O), Calc. 141.1279, Found 141.1281.

EXAMPLE 17 Preparation of (2S-cis)-3-Heptyloxymethyl)oxirane Methanol

The epoxidation was performed as described in Example 15 except that1.06 g (4.52 mmole) of L(+) diisopropyl tartrate, 0.92 g (3.23 mmole) ofTi(OiPr)₄, 6.35 ml (24.2 mmole) of 3.8 M tert-butylhydroperoxide inisooctane, and 3.00 g (16.1 mmole) of Z-4-(heptyloxy)-2-buten-1-ol wereused. The reaction yielded 2.76 g of white crystals followingcrystallization from Et₂ O: petroleum ether, mp 50-2° C. [∝]²⁵ -15.4' (c1.1, CHCl₃); ¹ H NMR (CHCl₃) δ 3.9-3.4 (m, 6), 3.25 (m, 2), 2.18 (dd, 1,J=6, 7 Hz), 1.60 (m, 2), 1.29 (br, 8), 0.88 (t, 3, J=6 Hz). Massspectra: EI; M-31 (--CH₂ OH), Calc. 171.1384, Found 171.1378.

EXAMPLE 18 Preparation of (2R-cis)-3-(Heptyloxymethyl)oxirane Methanol

The epoxidation was performed as described in Example 15 except thatD(-)diisopropyl tartrate was used. The reaction afforded 2.50 g of whitecrystals, mp 50-2° C. ²⁵ +13.2 (c, 1.2, CHCl₃); ¹ H NMR (CDCl₃), 3.9-3.4(m, 6), 3.25 (m, 2), 2.14 (dd, 1, J=6, 7 Hz), 1.60 (m, 2), 1.28 (br, 8),0.88 (t, 3, J=6 Hz). Mass spectra: EI, M-31 (--CH₂ OH), Calc. 171.1384,Found 171.1383.

EXAMPLE 19 Preparation of (2S*-cis)-3-Undecyloxiranecarboxylic Acid

A solution of 400 mg (1.89 mmole) of Z-2-tetradecen-1-ol and 13 mg (0.04mmole) of VO(acac)₂ in 50 ml of CH₂ Cl₂ was prepared under argon. Tothis solution was added 0.74 ml (2.83 mmole) of 3.8 M tert-butylhydroperoxide in isooctane. The mixture was allowed to stir for 2 daysunder argon. The mixture was poured into a chilled solution of 1.00 g ofFeSO₄ and 200 mg of citric acid in 10 ml H₂ O. The organic layer wasseparated and the aqueous layer washed twice with 10 ml of CH₂ Cl₂. Thecombined organic layers were dried over MgSO₄, filtered and evaporatedto afford a pale yellow solid. Flash chromatography (25 mm column; 50%Et₂ O; petroleum ether) yields 288 mg of the epoxy alcohol.

To a mixture of 3 ml CCl₄, 3 ml CH₃ CN, and 4.5 ml H₂ O was added 158 mg(0.44 mmole) of the epoxy alcohol followed by 5 mg of RuCl₃ --H₂ O. Themixture was stirred vigorously and 600 mg (2.63 mmole) of H₅ IO₆ wasadded in small portions over 20 min. The mixture was allowed to stirvigorously for three hours. The reaction was then diluted with 30 ml ofEt₂ O and the organic layer separated. The aqueous layer was washedtwice with 10 ml of Et₂ O. The combined organic layers were washed with20 ml of brine, dried over MgSO₄, filtered through a 2 cm pad of celiteand evaporated to afford a pale grey solid. Recrystallization from Et₂O: Petroleum ether yielded 106 mg of white solid, mp 82-3° C. ¹ H NMR δ3.6 (d, 1, J=5 Hz), 3.2 (m, 1), 1.7-1.2 (m, 20), 0.88 (t, 3, J=6 Hz).Mass Spectra: EI; M-45 (--CO₂ H), Calc. 197.1905, Found 197.1919.

EXAMPLE 20 Preparation of (2S*-trans)-3-Undecyloxiranecarboxylic Acid

A solution of 200 mg (0.95 mmole) of E-2-tetradecen-1-ol and 6 mg (0.02mmole) of VO(acac)₂ in 25 ml of CH₂ Cl₂ was prepared under argon. Tothis solution was added 0.38 ml (1.42 mmole) of 3.8 M tert-butylhydroperoxide in isooctane. The mixture was allowed to stir for 2 daysunder argon. The mixture was poured into a chilled solution of 0.50 g ofFeSO₄ and 100 mg of citric acid in 10 ml H₂ O. The organic layer wasseparated and the aqueous layer was washed twice with 10 ml of CH₂ Cl₂.The combined organic layers were dried over MgSO₄, filtered andevaporated to afford a pale yellow solid. Flash chromatography (25 mmcolumn; 50% Et₂ O;petroleum ether) yielded 128 mg of the epoxy alcohol.

To a mixture of 3 ml CCl₄, 3 ml CH₃ CN, and 4.5 ml H₂ O was added 100 mg(0.32 mmole) of the epoxy alcohol followed by 5 mg of RuCl₃ --H₂ O. Themixture was stirred vigorously and 400 mg (1.75 mmole) of H₅ IO₆ wasadded in small portions over 20 min. The mixture was allowed to stirvigorously for three hours. The reaction was then diluted with 30 ml ofEt₂ O and the organic layer separated. The aqueous layer was washedtwice with 10 ml of Et₂ O. The combined organic layers were washed with20 ml of brine, dried over MgSO₄, filtered through a 2 cm pad of celiteand evaporated to afford a pale grey solid. Recrystallization from Et₂O:Petroleum ether yielded 106 mg of white solid, mp 76-7° C., ¹ H NMR δ3.29 (d, 1, J=2 Hz), 3.2 (m, 1), 1.7-1.2 (m, 20), 0.88 (t, 3, J=6 Hz).Mass Spectra: EI, M-45 (--CO₂ H), Calc. 197.1905, Found 197.1919.

EXAMPLE 21 Preparation of (2R-cis)-3-Nonyloxiranecarboxylic Acid

Method A:

To a solution of 4 ml CH₃ CN, 4 ml CCl₄, and 6 ml H₂ O was added 20 mg(0.07 mmole) RuCl₃ --H₂ O and 400 mg (2.00 mmole) of(2S-cis)-nonyloxirane methanol. The solution was cooled to 0° C. and1.37 g (6.09 mmole) of H₅ IO₆ was added in small portions with vigorousstirring over 15 min. The mixture was stirred vigorously for one hour.The reaction was then poured into 30 ml of Et₂ O and the organic layerseparated. The organic layer was washed four times with 40 ml of 0.1 NNaOH. The-combined NaOH layers were acidified to pH 2 with conc. HCl andthe aqueous layer was washed four times with 30 ml of Et₂ O. Thecombined Et₂ O layers were washed with 30 ml of H₂ O, 30 ml of brine,dried over MgSO₄, filtered, and evaporated to give 340 mg of an offwhite solid. This residue was crystallized from Et₂ O: petroleum etherto afford 287 mg of white powder, mp 63-4° C. ¹ H NMR (CDCl₃) 3.58 (d,1, J=5 Hz), 3.24 (dd, 1, J=5, 12 Hz), 1.7-1.6 (m, 12), 0.88 (t, 3, J=6Hz); ²⁵ +9.5'. Mass Spectra: EI, M-45 (--CO₂ H), Calc. 169.1592, Found169.1596.

Method B:

To a solution of 500 mg (2.50 mmole) of (2S-cis)-3-nonyloxiranemethanolin 50 ml of DMF was added 3.30 g (8.75 mmole) of pyridinium dichromate.The reaction was allowed to stir overnight under argon. The mixture wasthen diluted with 150 ml H₂ O and 20 ml 0.1 M HCl and washed three timeswith 50 ml of Et₂ O. The combined Et₂ O layers are dried over MgSO₄,filtered and evaporated to afford a white powder. This residue whencrystallized from petroleum ether to afforded 268 mg of white powder, mp62-3° C.

EXAMPLE 22 Preparation of (2-S-cis)-3-Nonyloxiranecarboxylic Acid

The oxidation was performed as described in Example 21, Method A, exceptthat (2R-cis)-3-nonyloxirane methanol was used. The reaction affords 270mg of white solid, mp 63-4° C. ¹ H NMR (CDCl₃) 3.58 (d, 1, J=5 Hz), 3.24(dd, 1, J=5, 12 Hz), 1.7-1.6 (m, 12), 0.88 (t, 3, J=6 Hz). Mass Spectra:EI, M-45 (--CO₂ H), Calc. 169.1592, Found 169.1594.

EXAMPLE 23 Preparation of (2R-cis)-3-Nonyloxiranecarboxy Amide

Method A:

A solution of 50 mg of (2R-cis)-3-nonyloxirane carboxylic acid in 10 mlof Et₂ O was treated with a solution of CH₂ N₂ in Et₂ O (prepared fromN-nitrosomethylurea) at 0° C. until the yellow color persists for 20min. The mixture was allowed to stand for one hour and the Et₂ O wasevaporated under a stream of nitrogen. The residue was flashchromatographed (10 mm column; 40% Et₂ O;petroleum ether) to afford aclear oil.

A portion of the oil (25 mg) was dissolved in 3 ml of MeOH and 1 ml ofconc. NH₄ OH was added with stirring. The mixture was allowed to stirfor three days. The solution was then diluted with 15 ml of CHCl₃ andthe organic layer washed three times with 5 ml of 0.1 N HCl, once withhalf-saturated NaHCO₃, dried over MgSO₄, filtered and evaporated toyield a pale yellow oil. Flash chromatography (7 mm column; 5%MeOH:CHCl₃) afforded 11.2 mg of a clear oil which solidified onstanding. 1H NMR δ (CDCl₃) 6.09 (br, 1), 5.47 (br, 1), 3.49 (d, 1, J=5Hz), 3.19 (dd, 1, J=6, 11 Hz), 1.7-1.2 (m, 12), 0.88 (t, 3, J=6 Hz).Mass Spectra: EI; (M⁺) Calc. 213.1728, Found 213.1736:FAB; (M+H⁺) 214.

Method B:

A solution of 300 mg (1.40 mmole) of (2R-cis)-3-nonyloxiranecarboxylicacid and 0.235 ml (170 mg, 168 mmole) of triethylamine in 30 ml of THFwas prepared under argon. To this solution 210 mg (1.54 mmole) ofisobutylchloroformate was added and the mixture allowed to stir for 30min. Concentrated NH₄ OH (2 ml) was then added and the mixture stirredfor an additional 2 hours. The reaction was diluted with 75 ml of Et₂ O,washed with 50 ml of saturated NaHCO₃, 30 ml of brine, dried over MgSO₄,filtered and evaporated to yield a white residue. This residue was flashchromatographed (25 mm column; 5% MeOH:CHCl₃) to afford 268 mg of awhite solid, mp 91-2° C. ¹ H NMR was identical to the material preparedby Method A. ²⁵ +35.6'.

EXAMPLE 24 Preparation of (2S-cis)-3-Nonyloxiranecarboxy Amide

The amide was prepared as described in Example 23 except that(2S-cis)-3-nonyloxirane carboxylic acid was used. Method A afforded 16.6mg of a solid after chromatography. ¹ H NMR (CDCl₃) indicated a compoundwith an identical spectrum of that of the material prepared in example13 was present, contaminated with what appeared to be the parent acid.

Method B yielded 279 mg of white solid after chromatography, mp 85-8° C.¹ H NMR was the same as that of material prepared by method A. MassSpectra: FAB; (M+H⁺) 214.

EXAMPLE 25 Preparation of Methyl (2R-cis)-3-(benzyloxymethyl)oxiraneCarboxylate

To a solution of 10 ml CCl₄, 10 ml CH₃ CH, and 15 ml of H₂ O was added50 mg of RuCl₃ --H₂ O and 1.00 g (5.16 mmole) of(2S-cis)-3-(benzyloxymethyl)oxirane methanol. The solution was cooled to0° C. and 3.50 g (15.5 mmole) of H₅ IO₆ was added in portions over 15min with vigorous stirring. The reaction was allowed to stir vigorouslyfor 2 hours. The mixture was poured into 100 ml of Et₂ O, the organiclayer separated and the aqueous layer washed with 20 ml of Et₂ O. Thecombined organic layers were washed four times with 50 ml 0.1 N NaOH.The aqueous layers were combined, acidified to pH 2, and extracted threetimes with 50 ml Et₂ O. The ether layers were dried over MgSO₄, filteredand evaporated to yield a pale yellow residue.

The residue was dissolved in 40 ml of Et₂ O and treated with a solutionof CH₂ N₂ in Et₂ O until the yellow color persists for 30 min. Thesolution was allowed to stand for two hours. The Et₂ O layer wasevaporated under a stream of nitrogen and the residue flashchromatographed (25 mm column; 50% Et₂ O:petroleum ether) to afford 460mg of a clear oil. ¹ H NMR (CDCl₃) δ 7.4-7.2 (m, 5), 4.56 (dd, 2, J=12,22 Hz), 3.75 (S, 3), 3.73 (dd, 2, J=1, 5 Hz), 3.57 (d, 1, J=5 Hz), 3.44(m, 1). Mass Spectra: EI, M⁺ 222(2), 91(100), 107(81) FAB M+H⁺ 223.

EXAMPLE 26 Preparation of Methyl(2S-cis)-3-(benzyloxymethyl)oxiranecarboxylate

The compound was prepared as described in Example 25 except at twice thescale. After chromatography 985 mg of a clear oil was obtained. ¹ H NMR(CDCl₃) δ 7.4-7.2 (m, 5), 4.56 (dd, 2, J=12, 22 Hz), 3.75 (s, 3), 3.73(dd, 2, J-1, 5 Hz), 3.57 (d, 1, J=5 Hz), 3.44 (m, 1). Mass Spectra: EI,(M⁺) Calc. 222.0891, Found 222.0901.

EXAMPLE 27 Preparation of (2R-cis)-3-(Benzyloxymethyl)oxiraneCarboxamide

To a solution of 50 mg of methyl (2R-cis)-3-(benzyloxymethyl)oxiranecarboxylate in 3 ml of MeOH was added 1 ml of concentrated NH₄ OH withrapid stirring. The mixture was allowed to stir for two days. Thereaction was then diluted with 20 ml of CHCl₃ and the organic layerseparated and washed three times with 10 ml of 0.1 N HCl. The combinedorganic layers were dried over MgSO₄, filtered and evaporated to yield apale yellow residue. Flash chromatography (7 mm column, 5% MeOH: CHCl₃)afforded 24.2 mg of a clear oil which solidified upon standing. ¹ H NMR(CDCl₃) 7.4-7.2 (m, 5), 6.08 (br, 1), 5.64 (br, 1), 4.58 (dd, 2, J=12,15 Hz), 3.8 (m, 1), 3.5-3.4 (m, 3). Mass Spectra: EI: M-41 (--CONH₂)Calc. 163.0758, Found 163.0745.

EXAMPLE 28 Preparation of (2S-cis)-3-(benzyloxymethyl)oxiraneCarboxamide

The compound was prepared as described in Example 27 except that methyl(2S-cis)-3-(benzyloxymethyl)oxirane carboxylate was used. Afterchromatography 28.6 mg of clear oil was obtained which solidifies uponstanding. ¹ H NMR indicated the product was heavily contaminated withanother product, probably the acid. No further purification wasattempted. Mass Spectra: EI; M-44 (--CONH₂), Calc. 163.0758, Found163.0745.

EXAMPLE 29 Preparation of (2S-trans)-3-Nonyloxirane Methanol

A suspension of 1.00 g of 4a powdered molecular sieves in 50 ml of dryCH₂ Cl₂ was cooled to 0° C. under argon. To the cooled suspension wasadded 308 mg (1.09 mmole) of Ti(OiPr)₄ and 356 mg (1.52 mmole) ofL(+)diisopropyl tartrate via syringe. The mixture was stirred for 15 minand cooled to -20° C. A solution of tert-butyhydroperoxide in isooctane(3.8 M, 4.30 ml, 16.4 mmole) was added slowly to the mixture and thecatalyst was allowed to "age" for 30 min at -20° C. A solution of 2.00 g(10.9 mmole) of E-2-dodecen-1-ol in 10 ml of CH₂ Cl₂ was dried over 3Amolecular sieves for 15 ml before being added to the mixture slowly,with stirring, at -20°. The solution was then placed in the freezer(-30° C.) and allowed to stand for 24 H. The mixture was warmed to 0° C.and poured into a solution of 1.00 g of citric acid and 3.50 g of FeSO₄in 15 ml H₂ O. The organic layer was separated and the acqueous layerwashed twice with 20 ml of CH₂ Cl₂. The CH₂ Cl₂ layers were combined,dried over MgSO₄, filtered and evaporated to yield a sticky white solid.Recrystallization from Et₂ O:petroleum ether afforded 1.98 g of whitesolid, mp 62-4° C. ¹ H NMR δ 3.92 (ddd, 1, J=2.5, 6, 12 Hz), 3.63 (ddd,1, J=4, 7, 12 Hz), 2.95 (m, 2), 1.69 (dd, 1, J=7, 6 Hz), 1.6-1.2 (m,16), 0.88 (t, 3, J=6 Hz). Mass Spectra: M-31(--CH₂ OH)(0.4)169,M(100)55.1.

EXAMPLE 30 Preparation of (2R-trans)-3-Nonyloxirane Methanol

The epoxidation was performed as described of example 29 except thatD(-)diisopropyl tartrate was used. The reaction afforded 1.71 g. ofwhite solid, mp 57-60° C. ¹ H NMR (CDCl₃) δ 3.92 (ddd, 1, J=2.5, 6, 12Hz), 3.63 (ddd, 1, J=4, 7, 12 Hz), 2.95 (m, 2), 1.69 (dd, 1, J=7, 6 Hz),1.6-1.2 (m, 16), 0.88 (t, 3, J=6 hz). Mass Spectra: M-31(--CH₂OH)(0.6)169, M(100)55.1.

EXAMPLE 31 Preparation of (2S-trans)-3-Undecyloxirane Methanol

A suspension of 1.00 g of 4A powdered molecular sieves in 50 ml of dryCH₂ Cl₂ was cooled to 0° C. under argon. To the cooled suspension wasadded 403 mg (1.42 mmole) of Ti(OiPr)₄ and 463 mg (1.98 mmole) ofL(+)diisopropyl tartrate via syringe. The mixture was stirred for 15 minand cooled to -20° C. A solution of tert-butylhydroperoxide in isooctane(3.8 M, 5.58 ml, 21.2 mmole) was added slowly to the mixture and thecatalyst was allowed to "age" for 30 min at -20° C. A solution of 3.00 g(14.2 mmole) of E-2-tetradecen-1-ol in 10 ml of CH₂ Cl₂ was dried over3A molecular sieves for 15 min before being added to the mixture slowly,with stirring, at -20° C. The solution was then placed in the freezer(-30° C.) and allowed to stand of 24 H. The mixture was warmed to 0° C.and poured into a solution of 1.00 g of citric acid and 5.00 g of FeSO₄in 15 ml H₂ O. The organic layer was separated and the aqueous layerwashed twice with 20 ml of CH₂ Cl₂. The CH₂ Cl₂ layers are combined,dried over MgSO₄, filtered and evaporated to yield a sticky white solid.Recrystallization from Et₂ O:petroleum ether afforded 2.34 g of whitesolid, mp 68-70° C. ¹ H NMR δ 3.92 (ddd, 1, J=2.5, 6, 12 Hz), 3.63 (ddd,1, J=4, 7, 12 Hz), 2.95 (m, 2), 1.64 (dd, 1, J=7, 5 Hz), 1.6-1.2 (m,20), 0.88 (t, 3, J=5 Hz). Mass Spectra: M-31(--CH₂ OH)(1.0)197.2,M(100)55.1.

EXAMPLE 32 Preparation of (2R-trans)-3-Undecyloxirane Methanol

The epoxidation was performed as described of example 31 except thatD(-)diisopropyl tartrate was used. The reaction afforded 2.11 g of whitesolid, mp 66-8° C. ¹ H NMR δ 3.92 (ddd, 1, J=2.5, 6, 12 Hz), 3.63 (ddd,1, J=4, 7, 12 Hz), 2.95 (m, 2), 1.67 (dd, 1, J=7, 4 Hz), 1.6-1.2 (m,20), 0.88 (t, 3, J=5 Hz). Mass Spectra: M-31(--CH₂ OH)(1.0)197.2,M(110)55.1.

EXAMPLE 33 Preparation of (2S-trans)-3-heptyloxirane Methanol

A suspension of 2.00 g of 4A powdered molecular sieves in 50 ml of dryCH₂ Cl₂ was cooled to 0° C. under argon. To the cooled suspension wasadded 273 mg (1.00 mmole) of Ti(OiPr)₄ and 314 mg (1.34 mmole) ofL(+)diisopropyl tartrate via syringe. The mixture was stirred for 15 minand cooled to -20° C. A solution of tert-butylhydroperoxide in isooctane(3.8 M, 7.57 ml, 28.8 mmole) was added slowly to the mixture and thecatalyst was allowed to "age" for 30 min at -20° C. A solution of 3.00 g(19.2 mmole) of E-2-decen-1-ol in 10 ml of CH₂ Cl₂ was dried over 3Amolecular sieves for 15 min before being added to the mixture slowly,with stirring, at -20° C. The solution was then placed in the freezer(-30° C.) and allowed to stand for 24 H. The mixture was-warmed to 0° C.and poured into a solution of 1.00 g of citric acid and 5.00 g of FeSO₄in 15 ml H₂ O. The organic layer was separated and the aqueous layerwashed twice with 20 ml of CH₂ Cl₂. The CH₂ Cl₂ layers are combined,dried over MgSO₄, filtered and evaporated to yield a sticky white solid.Recrystallization from Et₂ O:petroleum ether afforded 2.43 g of whitesolid, mp 50-1° C. ¹ H NMR δ 3.92 (ddd, 1, J=2, 6, 12 Hz), 3.63 (ddd, 1,J=4, 8, 12 Hz), 2.94 (m, 2), 1.73 (dd, 1, J=6, 7 Hz), 1.6-1.2 (m, 12),0.88 (t, 3, J=7 Hz). Mass Spectra: M-31(--CH₂ OH)(0.5)148.1, M(100)69.1.

EXAMPLE 34 Preparation of (2R-trans)-3-Heptyloxirane Methanol

The epoxidation was performed as described of example 33 except thatD(-)diisopropyl tartrate was used. The reaction afforded 2.72 g of whitesolid, mp 48-50° C. ¹ H NMR (CDCl₃) δ 3.92 (ddd, 1, J=2, 6, 12 hz), 3.63(ddd, 1, J=4, 8, 12 Hz), 2.94 (m, 2), 1.70 (dd, 1, J=6, 7 hz), 1.6-1.2(m, 12), 0.88 (t, 3, J=7 Hz). Mass Spectra: M-31(--CH₂ OH)(0.5)141.1,M(100)69.1.

EXAMPLE 35 Preparation of (2R-trans)-3-Nonyloxiranecarboxylic Acid

To a solution of 2 ml CH₃ CN, 2 ml CCl₄, and 3 ml H₂ O was added 13 mg90.05 mmole) RuCl₃ --H₂ O and 500 mg 92.50 mmole) of(2S-trans)-nonyloxirane methanol. The solution was cooled to 0° C. and1.43 g (6.25 mmole) of H₅ IO₆ was added in small portions with vigorousstirring over 15 min. The mixture was stirred vigorously for one hour.The reaction was then poured into a mixture of 30 ml of Et₂ O and 10 mlof N HCl and the organic layer was separated. The aqueous layer waswashed three times with 30 ml of ether. The combined Et₂ O layers werewashed with 30 ml of H₂ O, 30 ml of brine, dried over MgSO₄, filtered,and evaporated to give 340 mg of an off white solid. The residue wasfiltered through a three inch pad of silica with ether and crystallizedfrom Et₂ O:petroleum ether to afford 293 mg of white powder, mp 56-7° C.¹ H NMR (CDCl₃) δ 3.27 (d, 1, J=2 hz), 3.19 (dt, 1, J=2, 6 Hz), 3.19(dt, 1, J=2, 6 Hz), 1.6-1.2 (m, 16), 0.88 (t, 3, J=6 Hz). Mass Spectra:M-45(--CO₂ H)(2.2)169.1, M(100)41.0.

EXAMPLE 36 Preparation of (2S-trans)-3-Nonyloxiranecarboxylic acid

Method A:

To a solution of 2 ml CH₃ CN, 2 ml CCl₄, and 3 ml H₂ O was added 13 mg(0.04 mmole) RuCl₃ --H₂ O and 400 mg (2.00 mole) of(2R-trans)-nonyloxirane methanol. The solution was cooled to 0° C. and1.14 g (5.00 mmole) of H₅ IO₆ was added in small portions with vigorousstirring over 15 min. The mixture was stirred vigorously for one hour.The reaction was then poured into 30 ml of Et₂ O and 10 ml of 1 n HCland the organic layer was separated. The aqueous layer was washed threetimes with 30 ml of ether. The combined Et₂ O layers were washed with 30ml of H₂ O, 30 ml of brine, dried over MgSO₄, filtered, and evaporatedto give 340 mg of an off white solid. This residue was filtered througha three inch pad of silica with other and crystallized from Et₂O:petroleum ether to afford 220 mg of white powder, mp 55-8° C. ¹ H NMR(CDCl₃) δ 3.27 (d, 1, J=2 Hz), 3.18 (dt, 1, J=2, 6 Hz), 1.6-1.2 (m, 16),0.88 (t, 3, J=6 Hz). Mass Spectra: M-45(--CO₂ H)(2.2)167.1, M(100)41.0.

Method B:

To a solution of 300 mg (1.50 mmole) of(2R-trans)-3-nonyloxiranemethanol in 50 ml of DMF was added 3.50 g (9.31mmole) of pyridinium dichromate. The reaction was allowed to stirovernight under argon. The mixture was then diluted with 150 ml H₂ O and20 ml 0.1 M HCl and washed three times with 50 ml of Et₂ O. The combinedEt₂ O layers are dried over MgSO₄, filtered and evaporated to afford awhite powder. This residue was crystallized from petroleum ether toafford 268 mg of white power, mp 62-3° C.

EXAMPLE 37 Preparation of (2R-cis)-3-Heptyloxirane Carboxylic Acid

To a solution of 300 mg (1.16 mmole) (2R-cis)-3-heptyloxiranemethanol in50 ml of DMF was added 2.62 g (6.97 mmole) of pyridinium dichromate. Thereaction was allowed to stir overnight under argon. The mixture was thendiluted with 150 ml H₂ O and 20 ml 0.1 M HCl and washed three times with50 ml of Et₂ O. The combined Et₂ O layers are dried over MgSO₄, filteredand evaporated to afford a white powder. This residue was crystallizedfrom petroleum ether to afford 155 mg of white power, mp 53-5° C. ¹ HNMR (CDCl₃) δ 3.60 (d, 1, J=5 Hz), 3.24 (dd, 1, J=5 12 Hz), 1.6-1.2 (m,12), 0.88 (t, 3, J=7 Hz). Mass Spectra: M-45(--CO₂ H)(3.7)141.1,M(100)41.0.

EXAMPLE 38 Preparation of (2S-cis)-3-Heptyloxiranecarboxylic Acid

To a solution of 500 mg (2.50 mmole) of (2R-cis)-3-heptyloxiranemethanolin 50 ml of DMF was added 3.30 g (8.75 mmole) of pyridinium dichromate.The reaction was allowed to stir overnight under argon. The mixture wasthen diluted with 150 ml H₂ O and 20 ml 0.1 M HCl and washed three timeswith 50 ml of Et₂ O. The combined Et₂ O layers were dried over MgSO₄,filtered and evaporated to afford a white powder. This residue wascrystallized from petroleum ether to afford 39 mg of white power, mp62-3° C. ¹ H NMR (CDCl₃) δ 3.59 (d, 1, J=5 Hz), 3.24 (m, 1), 1.6-1.2 (m,12), 0.88 (t, 3, J=7 Hz). Mass Spectra: M+(19.1)186.1, M-45(42.3)141.1,M(100)95.1.

EXAMPLE 39 Preparation of (2R-trans)-3-Undecyloxiranecarboxylic Acid

To a solution of 4 ml CH₃ CN, 4 ml CCl₄, and 6 ml H₂ O was added 20 mg(0.07 mmole RuCl₃ --H₂ O and 400 mg (2.00 mmole) of(2S-trans)-undecyloxirane methanol. The solution was cooled to 0° C. and1.37 g (6.09 mmole) of H₅ IO₆ was added in small portions with vigorousstirring over 15 min. The mixture was stirred vigorously for one hour.The reaction was then poured into 30 ml of Et₂ O and 10 ml of 1 H HCland the organic layer was separated. The aqueous layer was washed threetimes with 30 ml of ether. The combined Et₂ O layers were washed with 30ml of H₂ O, 30 ml of brine, dried over MgSO₄, filtered, and evaporatedto give an off-white solid. This residue was filtered through a threeinch pad of silica with ether and crystallized from Et₂ O:petroleumether to afford 240 mg of white powder, mp 66-8° C. ¹ H NMR (CDCl₃) δ3.27 (d, 1, J=2 Hz), 3.19 (dt, 1, J=2, 6 Hz), 1.6-1.2 (m, 20), 0.88 (t,3, J=7 Hz). Mass Spectra: M+(0.7)212.2, M-45(5.5)197.2, M(100)41.0.

EXAMPLE 40 Preparation of (2S-trans)-3-Undecyloxiranecarboxylic Acid

The reaction was performed as described for example 39 except that(2R-trans)-undecyloxirane methanol was used. After workup andcrystallization from Et₂ O:petroleum ether the reaction afforded 335 mgof white powder, mp 75-7° C. ¹ H NMR (CDCl₃) δ 3.27 (d, 1, J=2 Hz), 3.19(dt, 1, J=2, 6 Hz), 1.6-1.2 (m, 20), 0.88 (t, 3, J=7 Hz). Mass Spectra:M+(1.2)242.2, M-45(10.8)197.2, M(100)41.0.

EXAMPLE 41 Preparation of N-(2R-cis)-3-Nonyloxiraneacyl L-proline MethylEster

A solution of 50 mg (0.23 mmole) of (2R-cis)-3-nonyloxiranecarboxylicacid and 0.101 ml (71 mg, 0.70 mmole) of triethylamine in 5 ml of THFwas prepared under argon and cooled to 0° C. To this solution 33 mg(0.24 mmole) of isobutylchloroformate was added and the mixture wasallowed to stir for 30 min. L-Proline methyl ester hydrochloride (50 mg,0.30 mmole) was then added and the mixture was stirred for an additional2 hours. The reaction was diluted with 75 ml of Et₂ O, washed with 50 mlof saturated NaHCO₃, 30 ml of brine, dried over MgSO₄, filtered andevaporated to yield a white residue. This residue was flashchromatographed (25 mm column; 5% MeOH:CHCl₃) to afford 60 mg of a clearoil. ¹ H NMR reveals the oil to be a mixture of the cis- and trans-amideisomers (ca. 1:2 ratio). ¹ H NMR (trans) 4.55 (m, 1), 3.72 (s, 3), 3.58(d, 1, J=4); (cis) 4.82 (dd, 1, J=4, 8 Hz), 3.77 (s, 3), 3.52 (d, 1, J=4Hz);(cis and trans) 3.18 (m, 1), 2.4-1.8 (m, 4), 1.6-1.2 9m, 18), 0.88(t, 3, J=7 Hz). Mass Spectra: M+(0.5)325.2, M-CO₂ Me(8.7)266.2,M(100)70.1.

EXAMPLE 42 Preparation of N,N-Diethyl-(2R-cis)-3-nonyloxiranecarboxyAmide

A solution of 75 mg 90.35 mmole) of (2R-cis)-3-nonyloxiranecarboxylicacid and 0.148 ml (106 mg, 1.05 mmole) of triethylamine in 5 ml of THFwas prepared under argon and cooled to 0° C. To this solution 50 mg(0.37 mmole) of isobutylchloroformate was added and the mixture wasallowed to stir of 30 min. Diethylamine (0.109 ml, 76 mg, 1.05 mmole)was then added and the mixture stirred for an additional 2 hours. Thereaction was diluted with 75 ml of Et₂ O, washed with 50 ml of saturatedNaHCO₃, 30 ml of brine, dried over MgSO₄, filtered and evaporated toyield a clear residue. This residue was flash chromatographed (25 mmcolumn; 5% MeOH:CHCl₃) to afford 80 mg of a clear oil. ¹ H NMR δ 3.57(d, 1, J=4 Hz), 3.6-3.3 (m, 4), 3.2 (m, 1), 1.6-1.2 (m, 19), 1.13 (t, 3,J=7 Hz), 0.88 (t, 3, J=7 Hz). Mass Spectra: M+(1.5)269, M-OH(1.3)252,M(100)100.1.

EXAMPLE 43 Preparation of cis-2-Epoxydecene

A solution of 2.00 g (14.3 mmole) of cis-2-decene in 70 ml of CH₂ Cl₂was prepared and 3.40 g (15.7 mmole) of ca. 80% MCPBA was added in oneportion. The solution became slightly warm and was allowed to stirovernight. The m-chlorobenzoic acid which had precipitated was removedby filtration and the filtrate was washed twice with 100 ml of 1 N NaOH.The organic layer was dried over MgSO₄, filtered and the solventevaporated to give a clear oil. After kugelrohr distillation (50-60° C.oven, 0.2 mm) 1.836 g of a clear oil was obtained. ¹ H NMR δ 3.04 (ddd,1, J=4, 5, 10), 2.90 (m, 1), 1.6-1.1 (m, 15), 0.88 (t, 3, J=7 Hz).

In-vitro Screening Anti--HIV XTT Assay

This assay, which is available to the public by the National CancerInstitute, utilizes colorimetry based on the production of a coloredformazin from a tetrazolium salt by viable cells, to develop a safe,rapid and quantitative measure of HIV cytopathology. The assay is amodification of the previously described screening method for thedetection of anti-tumor drug cytotoxicity (D. A. Scudiero, et al, CancerRes, 48, 4827-4833 (1988)). It involves the plating of susceptible human"host" cells with and without virus in microculture plates, addingvarious concentrations of the respective test compounds, incubating theplates for seven days, during which time infected, non-drug treatedcontrol cells are largely or totally destroyed by the virus. The numberof remaining viable cells are determined utilizing a calorimetricendpoint. The results are summarized in Table 1 which appears below. Abeneficial antiviral effect is indicated by a response % grater than thevirus kill %, with 100% being optimal.

                  TABLE 1                                                         ______________________________________                                        Compound        Dose     Antiviral                                                                             Virus  Virus                                 of Example                                                                            RUN     ug/ml    Effect  Source Kill                                  ______________________________________                                        11      1       10       60.2%   C      6.0%                                          2       1        37.5%   C      25.0%                                         3       23       19.9%   C      14.0%                                         4       11.5     18.1%   C      12.5%                                         5       31.9     23.9%   C      13.0%                                         6       11.5     12.3%   C      9.0%                                          7       31.9       13%   C      11.0%                                         8       10        110%   V      25.0%                                         9       23         30%   V      23.0%                                         10      11.5     68.5%   V      49.0%                                         11      31.9     65.1%   V      53.0%                                         12      11.5     80.4%   V      30.0%                                         13      31.9     82.6%   V      22.0%                                         14      3.9      21.5%   V      20.0%                                         15*     3.9      32.4%   V      25.0%                                 21      1       1        18.9%   C      9.0%                                          2       10       27.3%   C      22.0%                                         3       6.3      16.6%   C      12.0%                                         4       31.5     19.6%   C      14.0%                                         5       24.6     21.6%   C      14.0%                                         6       31.5     10.7%   C      12.0%                                         7       24.6       13%   C      18.0%                                         8       10         94%   V      23.0%                                         9       6.3      25.7%   V      19.0%                                         10      31.5     75.3%   V      53.0%                                         11      24.6     64.3%   V      57.0%                                         12      31.5       62%   V      27.0%                                         13      24.6       88%   V      40.0%                                         14      27.2     25.7%   V      18.0%                                         15*     27.2     28.1%   V      25.0%                                 23      1       28       75.6%   C      6.0%                                          2       76       28.5%   C      17.0%                                         3       38       35.1%   C      14.0%                                         4       140      25.1%   C      14.0%                                         5       38         17%   C      17.0%                                         6       14         16%   C      17.0%                                         7       2.8       108%   V      19.0%                                         8       7.6      40.2%   V      30.0%                                         9       3.8      60.1%   V      52.0%                                         10      14       60.1%   V      52.0%                                         11      380        24%   V      20.0%                                         12      140        31%   V      20.0%                                         13*     27       79.6%   V      22.0%                                 12      1       10       75.1%   C      6.0%                                          2       10       71.3%   C      27.0%                                         3       50       32.1%   C      16.0%                                         4       4.9      31.6%   C      16.0%                                         5       2.9      23.1%   C      15.0%                                         6       4.9      16.7%   C      11.0%                                         7       29       15.7%   C      11.0%                                         8       50       56.9%   V      27.0%                                         9       4.9      82.4%   V      53.0%                                         10      2.9      82.9%   V      55.0%                                         11      49       54.9%   V      30.0%                                         12      2.9      62.2%   V      36.0%                                         13      21       53.4%   V      19.5%                                         14      21       98.7%   V      24.0%                                 22      1       10       50.1%   C      21.0%                                         2       0.1      25.6%   C      20.0%                                         3       0.1       9.5%   C      8.0%                                          4       34.6     25.9%   C      16.0%                                         5       29       18.5%   C      13.0%                                         6       34.6     12.8%   C      14.0%                                         7       2.9      11.5%   C      16.0%                                         8       34.6     85.5%   V      54.0%                                         9       29       68.5%   V      55.0%                                         10      34.6     40.3%   V      26.0%                                         11      29       41.9%   V      33.0%                                         12      24.8     23.1%   V      30.0%                                         13*     24.8     26.6%   V      20.0%                                 24      1       35.2       73%   C      16.0%                                         2       20.7     67.4%   C      16.0%                                         3       3.52     11.3%   C      9.0%                                          4       2.07     10.5%   C      9.0%                                          5       35.2       73%   V      54.6%                                         6       20.7     71.4%   V      54.6%                                         7       3.52     28.1%   V      26.0%                                         8       20.7       34%   V      26.0%                                         9       1.34     41.7%   V      20.0%                                         10*     0.135     114%   V      23.0%                                 13      1       0.1      22.1%   C      21.0%                                         2       1        15.7%   C      13.0%                                         3       1        10.2%   C      11.0%                                         4       0.1        50%   V      55.0%                                         5       0.1      52.5%   V      27.0%                                         6       4.4      28.4%   V      24.0%                                          7*     0.44     30.1%   V      27.0%                                 25      1       100      28.6%   C      20.0%                                         2       100      19.5%   C      13.0%                                         3       10       12.3%   C      11.0%                                         4       10       46.5%   V      55.0%                                         5       1          49%   V      27.0%                                         6       4.7      25.6%   V      22.0%                                          7*     4.7      28.5%   V      22.0%                                 14      1       0.1      21.9%   C      18.0%                                         2       10       53.5%   C      17.0%                                         3       10         61%   C      17.0%                                         4       0.1      12.1%   C      10.0%                                         5       10       10.5%   C      10.0%                                         6       1        43.5%   V      50.0%                                         7       10       49.5%   V      50.0%                                         8       0.1      17.7%   V      16.0%                                         9       0.1      20.6%   V      16.0%                                         10      4.1      28.8%   V      29.0%                                         11*     0.004      30%   V      25.0%                                 26      1       10       20.8%   C      17.0%                                         2       10       39.4%   C      13.2%                                         3       100        54%   C      13.2%                                         4       10       13.8%   C      12.0%                                         5       100      16.3%   C      12.0%                                         6       10       58.7%   V      54.0%                                         7       100      81.3%   V      54.0%                                         8       1        19.8%   V      23.0%                                         9       100      23.7%   V      23.0%                                         10      487      28.1%   V      30.0%                                         11*     0.487    26.6%   V      25.0%                                 19      1       1        22.6%   C      21.0%                                         2       10       55.7%   C      15.0%                                         3       1        11.6%   C      13.0%                                         4       10       63.9%   V      57.0%                                         5       10       38.8%   V      26.0%                                         6       0.1      27.6%   V      19.0%                                          7*     1        12.5%   V      25.0%                                 20      1       10       22.6%   C      18.0%                                         2       10       59.5%   C      14.0%                                         3       100      60.9%   C      14.0%                                         4       100        10%   C      10.0%                                         5       100      10.2%   C      10.0%                                         6       100      75.7%   V      55.0%                                         7       100      84.3%   V      55.0%                                         8       100      40.2%   V      24.0%                                         9       100      72.5%   V      24.0%                                         10      4.07     45.5%   V      25.0%                                         11*     4.07     87.4%   V      23.0%                                 15      1       46       58.3%   C      15.0%                                         2       4.6      11.8%   C      13.0%                                         3       92       60.8%   V      35.0%                                         4       10       78.2%   V      57.0%                                         5       10       58.6%   V      26.0%                                          6*     21.6       32%   V      31.0%                                 16      1       10.6     27.3%   C      14.0%                                         2       80.2     69.6%   C      15.0%                                         3       80.2     84.5%   C      15.0%                                         4       160      62.6%   V      40.0%                                         5       80.2     97.3%   V      60.0%                                         6       80.2     88.6%   V      30.0%                                         7       23.8     35.9%   V      22.0%                                          8*     23.8     39.7%   V      28.0%                                 17      1       113      31.1%   C      10.0%                                         2       56.7     53.7%   C      15.0%                                         3       56.7     83.1%   C      15.0%                                         4       11.3     57.6%   V      40.0%                                         5       56.7      102%   V      57.0%                                         6       56.7     69.2%   V      26.0%                                          7*     1.97     15.8%   V      28.0%                                 18      1       96.5     38.6%   C      11.0%                                         2       48.4     30.7%   C      16.0%                                         3       4.84       17%   C      10.0%                                         4       9.65     58.8%   V      40.0%                                         5       48.4     89.8%   V      63.0%                                         6       48.4       49%   V      26.0%                                         7       2.4      29.4%   V      20.0%                                          8*     2.4      40.2%   V      25.0%                                 ______________________________________                                         Anti-viral effect: Response (Viable Cells, determined by colorometric         absorbance of formazan dye produced) as % of uninfected controls              Virus Source: HIV1 IIIb; C, Virus producing H9 cells; V, cell free virus      Virus Kill: Infected untreated cells as % of uninfected cell controls         *: Culture redosed with compound after 3 days                            

Activity Against HIV Protease

The assay for proteolyic activity is a modified version of thatpreviously described by Copeland et al., Gene Anal. Tech., 5:109-115(1988). For each assay to be performed 2 microliters of proteasesolution (ca. 100 ng) is added to 3 μg of nonapeptide(Val-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-NH₂) in 8 μl of 200 mM sodiumphosphate, 1 M NaCl, 5% glycerol, 0.25% Nonidet-40, at pH 6.5. Thisaffords a stock solution containing 10 μl per number of reactions. Thisstock is aliquoted out in 10 μl portions to which 0.5 μl of DMSO (acontrol is run with each set of assays to compensate for minorexperimental variations) or a DMSO solution containing 10 μg/μl of aninhibitor, i.e., any one of the compounds of the present invention, isadded. The reactions were incubated at room temperature and 4 μlportions were removed at ca. 16 and 40 hr. intervals. HPLC analysis wasperformed on a 4.6×250 mm Lichrosorb RP-18 column eluted with a gradientof 0-40% acetonitrile in water (containing 0.05% TFA) over 30 minmonitoring absorbance at 206 nm. Progress of the enzymatic cleavage wasfollowed by monitoring the appearance of the pentapeptide(Val-Ser-Gln-Asn-Tyr-OH) cleavage product. The activity is expressed asthe amount of the pentapeptide (Val-Ser-Gln-Asn-Tyr-OH) observedrelative to the control reaction for that set of assays. The results arereported in Table 2 below.

                  TABLE 2                                                         ______________________________________                                        Inhibition of HIV-2 protease                                                                            Percent Control                                     Ex-                       Cleavage of                                         ample                     Nonapeptide HIV-1                                   #     Compound            Cleavage Site Fragment                              ______________________________________                                              Myristic Acid       111%                                                      Cerulenin            10%                                                      Pepstatin A          1%                                                       1,2-Epoxy-3-(p-      18%                                                      nitrophenoxy)propane                                                     3    cis-2-Decenol       105%                                                12    2R-cis-Nonyloxirane methanol                                                                       96%                                                11    2S-cis-Nonyloxirane methanol                                                                       85%                                                16    2R-cis-Heptyloxirane methanol                                                                      42%                                                15    2S-cis-Heptyloxirane methanol                                                                      59%                                                18    2R-cis-(Heptyloxymethyl) oxirane                                                                   71%                                                      methanol                                                                17    2S-cis-(Heptyloxymethyl) oxirane                                                                   66%                                                      methanol                                                                19    2-cis-Undecyloxirane methanol                                                                     104%                                                14    2R-cis-(Benzyloxymethyl) oxirane                                                                   78%                                                      methanol                                                                13    2S-cis-(Benzyloxymethyl) oxirane                                                                   81%                                                      methanol                                                                43    cis-2-Epoxydecene   102%                                                30    2R-trans-Nonyloxirane methanol                                                                    105%                                                29    2S-trans-Nonyloxirane methanol                                                                    106%                                                34    2R-trans-Heptyloxirane methanol                                                                   104%                                                33    2S-trans-Heptyloxirane methanol                                                                   105%                                                32    2R-trans-Undecyloxirane methanol                                                                   98%                                                31    2S-trans-Undecyloxirane methanol                                                                   95%                                                20    2-trans-Undecyloxirane methanol                                                                    88%                                                21    2R-cis-Nonyloxiranecarboxylic acid                                                                 56%                                                22    2S-cis-Nonyloxiranecarboxylic acid                                                                 60%                                                37    2R-cis-Heptyloxiranecarboxylic acid                                                               104%                                                38    2S-cis-Heptyloxiranecarboxylic acid                                                               105%                                                19    2-cis-Undecyloxiranecarboxylic acid                                                                77%                                                35    2R-trans-Nonyloxiranecarboxylic                                                                    77%                                                      acid                                                                    36    2S-trans-Nonyloxiranecarboxylic                                                                    12%                                                      acid                                                                    39    2R-trans-Undecyloxiranecarboxylic                                                                  84%                                                      acid                                                                    40    2S-trans-Undecyloxirane                                                                            93%                                                      carboxylic acid                                                         23    2R-cis-Nonyloxiranecarboxy amide                                                                  102%                                                24    2S-cis-Nonyloxiranecarboxy amide                                                                  113%                                                      Cerulenin            10%                                                42    N,N-Diethyl-2R-Cis-  77%                                                      nonloxiranecarboxy amide                                                41    N-(2R-cis-Nonyloxiraneacyl)-L-                                                                     24%                                                      proline methyl ester                                                    ______________________________________                                    

Renin Activity: Porcine Renin Cleavage of Porcine Tetradecapeptide

Proteolytic enzymes are ubiquitous in many living systems. Renin is anasparxyl protease found in mammals which is responsible for theregulation of blood pressure. Structural and functional similaritiesbetween Renin and retroviral proteases have been previously noted(Pearl, L. H., Taylor, W. R., (1987) Nature, 329, 351-354).Consequently, successful therapeutic inhibition of retroviral proteasesdepends on the ability of the inhibitor to inhibit the retroviral enzymewhile having a greatly diminished effect on normally occurring enzymeslike Renin. The failure to inhibit the desired enzyme selectively mayhave an adverse effect on normal biochemical functions. The similarityof Renin to retroviral proteases and its viral normal bodily functionmakes it an attractive system in which to examine inhibitoryselectivity.

Renin activity was determined by a method similar to that describedabove for the HIV protease. A solution containing 0.37 m unit (0.5 μl ofstock solution containing 0.75 m unit/μl) of porcine Renin (Sigma) wasadded to 5 μg of porcine angiotensinogen in 50 μl of 0.1 Mcitrate-phosphate buffer (pH 6.00) containing 0.25% NP-40. A DMSOsolution containing the inhibitor (0.5 μl of a 50 μg/μl solution) wasadded and the reaction was allowed to incubate at room temperature for70 min. The entire mixture was analyzed by HPLC chromatographic analysiswas perforemd on a 4.6×250 mm Lichrosorb RP-18 column eluting with agradient of 0-50% acetonitrile in 0.02 M KH₂ Po₄ (pH 4.7) over 30 minmonitoring absorbance at 206 nm. Appearance of both the tetrapeptide andthe decapeptide products may be monitored under these conditions. Enzymeactivity is expressed as the amount of tetrapeptide produced relative toan uninhibited control, and are summarized in Table 3 below.

                  TABLE 3                                                         ______________________________________                                        Compound                      Percent control                                 of                            (tetrapeptide                                   Example Compound              produced)                                       ______________________________________                                        35      2R-trans-Nonyloxiranecarboxylic acid                                                                72%                                             36      2S-trans-Nonyloxiranecarboxylic acid                                                                12%                                             39      2R-trans-Undecyloxiranecarboxylic acid                                                              51%                                             40      2S-trans-Unceyloxiranecarboxylic acid                                                               71%                                             21      2R-cis-Nonyloxiranecarboxylic acid                                                                  86%                                             15      2S-cis-Heptyloxirane methanol                                                                       86%                                             16      2R-cis-Heptyloxirane methanol                                                                       96%                                             11      2S-cis-Nonyloxirane methanol                                                                        106%                                            23      2R-cis-Nonyloxiranecarboxy amide                                                                    88%                                             24      2S-cis-Nonyloxiranecarboxy amide                                                                    106%                                                    Cerulenin             68%                                                     Epoxy-3-(p-nitrophenoxy) propene                                                                    106%                                                    Pepstatin A                                                           ______________________________________                                    

The compounds of the present invention may be made into pharmaceuticalcompositions by combination with appropriate pharmaceutically acceptablecarriers or diluents, and may be formulated into preparations in solid,semisolid, liquid or gaseous forms such as tablets, capsules, powders,granules, ointments, solutions, suppositories, injections, inhalants,and aerosols in the usual ways for their respective route ofadministration. The following methods and excipients are merelyexemplary and are in no way limiting.

In pharmaceutical dosage forms, the compounds of the present inventionmay be used in the form of their pharmaceutically acceptable salts, andalso may be used alone or in appropriate association, as well as incombination with other pharmaceutically active compounds.

In the case of oral preparations, the compounds may be used alone or incombination with appropriate additives to make tablets, powders,granules or capsules, e.g., with conventional additives such as lactose,mannitol, corn starch or potato starch; with binders such as crystallinecellulose, cellulose derivatives, acacia, corn starch or gelatins; withdisintegrators such as corn starch, potato starch or sodiumcarboxymethyl-cellulose; with lubricants such as talc or magnesiumstearate; and if desired, with diluents, buffering agents, moisteningagents, preservatives and flavoring agents.

Furthermore, the compounds of the present invention may be made intosuppositories by mixing with a variety of bases such as emulsifyingbases or water-soluble bases.

The compounds of the present invention may be formulated intopreparations for injections by dissolving, suspending or emulsifyingthem in an aqueous or non-aqueous solvent, such as vegetable oil,synthetic aliphatic acid glycerides, esters of higher aliphatic acids orpropylene glycol; and if desired, with conventional additives such assolubilizers, isotonic agents, suspending agents, emulsifying agents,stabilizers and preservatives.

In the cases of inhalations or aerosol preparations, the compounds ofthe invention in the form of a liquid or minute powder may be filled upin an aerosol container with gas or liquid spraying agents, and ifdesired, together with conventional adjuvants such as humidifyingagents. They may also be formulated as pharmaceuticals fornon-pressurized preparations such as in a nebulizer or an atomizer.

The amount of the compounds of the present invention to be used variesaccording to the degree of the infection encountered, and the stages ofthe disease. A suitable dosage is about 0.5 to 100 mg/kg body weight.The preferred dosage is that amount sufficient to render a hostasymptomatic to the particular viral infection. The dose may vary whenthe compounds are used prophylactically.

A method of treatment of retroviral infections utilizing the 2,3-epoxycompounds of the present invention can generally be by oral ingestionwith a pharmaceutically acceptable carrier. The 2,3-epoxy compounds ofthe present invention can also be administered systemically, e.g.,parenterally, via inhalation, or rectally to a person infected by retrovirus.

Unit dosage forms for oral administration such as syrups, elixirs, andsuspensions wherein each dosage unit, e.g., teaspoonful, tablespoonful,contains a predetermined amount of the 2,3-epoxy compound of the presentinvention. Inclusion of pharmaceutically acceptable excipients, arereadily known by those skilled in the art. Parenteral administration ofthe 2,3-epoxy compounds of the present invention can be by apharmaceutically acceptable carrier, such as Sterile Water forInjection, USP, or by normal saline.

The 2,3-epoxy compounds of the present invention can be administeredrectally via a suppository. The suppository can include vehicles such ascocoa butter, carbowaxes and polyethylene glycols, which melt at bodytemperature, yet are solidified at room temperature.

The 2,3-epoxy compound of the present invention can be utilized inaerosol formulation to be administered via inhalation. The 2,3-epoxycompounds can be formulated into pressurized aerosol containers togetherwith a pharmaceutically acceptable propellants such asdichlorodifluoromethane, propane, nitrogen and the like.

The term "unit dosage form" as used herein refers to physically discreteunits suitable as unitary dosages for human and animal subjects, eachunit containing a predetermined quantity of the 2,3-epoxy compoundcalculated in an amount sufficient to produce the desired to effect inassociation with a pharmaceutically acceptable, diluent, carrier orvehicle. The specifications for the novel unit dosage forms of thepresent invention depend on the particular compound employed and theeffect to be achieved, and the pharmacodynamics associated with eachcompound in the host.

The pharmaceutically acceptable adjuvants, for example, vehicles,carrier of diluents are readily available to the public. The amount ofthe anti-retroviral 2,3-epoxy compound suitable for the various dosageforms can be determined by the particular anti-retroviral activity ofeach compound per se.

While not being bound to any theory, it is believed that the 2,3-epoxycompounds inhibit the essential protein enzyme necessary for viralreplication. The details of the assay are set forth further in Copelandet al., Genetic Locus, Primary Structure, and Chemical Synthesis ofHuman Immunodeficiency Virus Protease, Gene Anal Techn 5:109-115 (1988).

Any necessary adjustments in dose can be readily made to meet theseverity of the infection and adjusted accordingly by the skilledpractitioner.

We claim:
 1. An optical isomer of an anti-viral compound of the formula:##STR7## wherein R is selected from --CO₂ R², --CONR³ R⁴ or COR⁵,wherein R³ and R⁴ are each independently hydrogen or a lower alkylgroup, R⁵ is a peptide having at least two amino acid residues;andwherein R¹ is aralkyl with the alkyl group having 1-6 carbon atoms,aralkyl (lower alkyl) ether with the alkyl group having 1 to 4 carbonatoms or C₅ -C₁₃ alkyl (lower alkyl) ether.
 2. An optical isomer of ananti-viral compound of the formula: ##STR8## wherein R⁵ is a peptidehaving at least two amino acid residues; and wherein R¹ is C₅ -C₁₃alkyl.
 3. An anti-viral composition comprising an effective anti-viralamount of an optical isomer of a compound having the formula: ##STR9##wherein R is selected from --CH₂ OH, --CONR³ R⁴, or COR⁵, wherein R³ andR⁴ are each independently hydrogen or a lower alkyl group, R⁵ is apeptide having at least two amino acid residues;and wherein R¹ isaralkyl, aralkyl (lower alkyl) ether or C₅ -C₁₃ alkyl (lower alkyl)ether; and a pharmaceutically acceptable carrier.
 4. An anti-viralcomposition comprising an effective anti-viral amount of an opticalisomer of an a compound having the formula: ##STR10## wherein R⁵ is apeptide having at least two amino acid residues; wherein R¹ is C₅ -C₁₃alkyl: anda pharmaceutically acceptable carrier.
 5. A method of treatingviral infections to a host in need thereof by administering an effectiveanti-viral amount of an optical isomer of the compound having theformula: ##STR11## wherein R is selected from --CH₂ OH, --CO₂ R²,--CONR³ R⁴, or COR⁵, wherein R² is hydrogen or a lower alkyl group, R³and R⁴ are each independently hydrogen or a lower alkyl group, R⁵ is apeptide having at least two amino acid residues; andwherein R¹ is C₅-C₁₃ alkyl, aryl, aralkyl, aralkyl (lower alkyl) ether, or C₅ -C₁₃ alkyl(lower alkyl) ether.
 6. The method according to claim 5 wherein R is--COOH.
 7. The method according to claim 5 wherein R is --CH₂ OH.
 8. Themethod according to claim 6 wherein said compound is2-S-trans-nonyloxirane carboxylic acid.
 9. The method according to claim5 wherein R and R¹ are cis to each other.
 10. The method according toclaim 5 wherein R and R¹ are trans to each other.
 11. The method ofclaim 5 wherein said viral infection is caused by a retrovirus.
 12. Themethod according to claim 11 wherein said retrovirus is HIV-I.
 13. Themethod according to claim 11 wherein said retrovirus is HIV-II.